LOCUS       Exported                6301 bp ds-DNA     circular SYN 08-APR-2018
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6301)
  AUTHORS   LeonardLab
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 10, 2020 from SnapGene Viewer 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6301
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             910..975
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             982..1002
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1027..1176
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             1207..1989
                     /codon_start=1
                     /product="From Chavez et al., Nature Methods 2015"
                     /label=p65 AD
                     /translation="SQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRR
                     IAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQ
                     APAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDE
                     DLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGA
                     QRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALL"
     CDS             2008..2577
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     CDS             2617..2889
                     /codon_start=1
                     /product="From Khalil et al., Cell 2012"
                     /label=ZF97-4
                     /translation="ARPGERPFQCRICMRNFSRQSNLSRHTRTHTGEKPFQCRICMRNF
                     SRNEHLVLHLRTHTGEKPFQCRICMRNFSQKTGLRVHLKTHLRGS"
     polyA_signal    2940..3164
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3210..3638
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      3708..3843
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(4549..5134)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5305..6165)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6166..6270)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagtcccaa gctggctagc
      901 gccaccatgg attataagga tcacgatggc gactacaagg accacgatat tgactacaaa
      961 gacgatgacg ataaaatggc tcctaagaaa aagcgcaaag tcggtatcca tggcgttccc
     1021 ggatcagacg cattggacga ttttgatctg gatatgctgg gaagtgacgc cctcgatgat
     1081 tttgaccttg acatgcttgg ttcggatgcc cttgatgact ttgacctcga catgctcggc
     1141 agtgacgccc ttgatgattt cgacctggac atgctgatta actctagaag ttccggatct
     1201 gttggtagcc agtacctgcc cgacaccgac gaccggcacc ggatcgagga aaagcggaag
     1261 cggacctacg agacattcaa gagcatcatg aagaagtccc ccttcagcgg ccccaccgac
     1321 cctagacctc cacctagaag aatcgccgtg cccagcagat ccagcgccag cgtgccaaaa
     1381 cctgcccccc agccttaccc cttcaccagc agcctgagca ccatcaacta cgacgagttc
     1441 cctaccatgg tgttccccag cggccagatc tctcaggcct ctgctctggc tccagcccct
     1501 cctcaggtgc tgcctcaggc tcctgctcct gcaccagctc cagccatggt gtctgcactg
     1561 gctcaggcac cagcacccgt gcctgtgctg gctcctggac ctccacaggc tgtggctcca
     1621 ccagccccta aacctacaca ggccggcgag ggcacactgt ctgaagctct gctgcagctg
     1681 cagttcgacg acgaggatct gggagccctg ctgggaaaca gcaccgatcc tgccgtgttc
     1741 accgacctgg ccagcgtgga caacagcgag ttccagcagc tgctgaacca gggcatccct
     1801 gtggcccctc acaccaccga gcccatgctg atggaatacc ccgaggccat cacccggctc
     1861 gtgacaggcg ctcagaggcc tcctgatcca gctcctgccc ctctgggagc accaggcctg
     1921 cctaatggac tgctgtctgg cgacgaggac ttcagctcta tcgccgatat ggatttctca
     1981 gccttgctgg gctctggcag cggcagccgg gattccaggg aagggatgtt tttgccgaag
     2041 cctgaggccg gctccgctat tagtgacgtg tttgagggcc gcgaggtgtg ccagccaaaa
     2101 cgaatccggc catttcatcc tccaggaagt ccatgggcca accgcccact ccccgccagc
     2161 ctcgcaccaa caccaaccgg tccagtacat gagccagtcg ggtcactgac cccggcacca
     2221 gtccctcagc cactggatcc agcgcccgca gtgactcccg aggccagtca cctgttggag
     2281 gatcccgatg aagagacgag ccaggctgtc aaagcccttc gggagatggc cgatactgtg
     2341 attccccaga aggaagaggc tgcaatctgt ggccaaatgg acctttccca tccgccccca
     2401 aggggccatc tggatgagct gacaaccaca cttgagtcca tgaccgagga tctgaacctg
     2461 gactcacccc tgaccccgga attgaacgag attctggata ccttcctgaa cgacgagtgc
     2521 ctcttgcatg ccatgcatat cagcacagga ctgtccatct tcgacacatc tctgttttct
     2581 ggttctggtt ctggttcagg atccggtacc ggtggagcta gacccgggga gcgccccttc
     2641 cagtgtcgca tttgcatgcg gaacttttcg agacaatcta atttgtcaag acatacccgt
     2701 actcataccg gtgaaaaacc gtttcagtgt cggatctgta tgcgaaattt ctccagaaat
     2761 gaacatttgg ttttgcatct acgtacgcac accggcgaga agccattcca atgccgaata
     2821 tgcatgcgca acttcagtca aaaaactggt ttgagagttc acctaaaaac ccacctgaga
     2881 ggatcttaag cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
     2941 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3001 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3061 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3121 ggaagtcaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3181 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3241 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3301 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3361 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3421 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3481 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3541 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3601 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3661 ttaggctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3721 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3781 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3841 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga ggttgggctt
     3901 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     3961 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4021 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4081 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     4141 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4201 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4261 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4321 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4381 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4441 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4501 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4561 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4621 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4681 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4741 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4801 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4861 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4921 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4981 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5041 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa
     5101 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5161 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5221 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5281 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5341 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5401 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagatc cacgctcacc
     5461 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5521 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5581 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5641 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5701 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5761 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5821 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5881 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5941 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6001 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6061 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6121 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6181 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6241 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6301 c
//
